Bakery products comprising carbohydrate oxidase and/or pyranose oxidase

ABSTRACT

The present invention is related to a method for the improvement of shape and width of cuts of bread during the baking process of bakery products which comprises the step of adding a sufficiently effective amount of Carbohydrate oxidase or Pyranose oxidase in said bakery products.

FIELD OF THE INVENTION

The present invention is related to improvement of the shape of bread byaddition of functional ingredient such as enzymes.

STATE OF THE ART

Bread-improving and/or dough improving additives are in the bread makingprocess added to the dough in order to improve texture, volume, flavourand freshness of the bread as well as to improve mach inability andstability of the dough. Dough conditioners: Data—esters, oxidants likeAscorbic acid, KBrO₃, ADA and enzymes like lipase added to strengthenthe gluten and improve rheological and handling properties of the dough,improve the shape of the bread significantly. Dough pieces of baguettetype breads and schnittbrötchen are cut respectively after proofing,right before baking, and before proofing in order to improve the shapeof the final product due to increased opening of the cuts as a result ofthe improved oven spring.

As the use of chemical oxidants like ADA or bromate is forbidden in mostcountries research is performed in order to replace the chemicaloxidants by natural ones like enzymes.

Use of Glucose Oxidase, Carbohydrate Oxidase and Pyranose Oxidase inBaking

The effect of Carbohydrate oxidase and Pyranose oxidase as a doughimproving and bread improving agent are disclosed in respectively WO99/31990 and WO 97/22257.

EP 321 811 and EP 338 452 disclose the use of Glucose oxidase, incombination with other enzymes.

The inventors of WO99/31990 found a novel Carbohydrate oxidase which canoxidize the reducing end of an oligosaccharide more efficiently than thecorresponding monosaccharide, e.g., preferentially oxidizingmaltodextrines or cellodextrines over glucose. This document describesthe effect of carbohydrate oxidase on firmness, stickiness, stabilityand robustness of the dough. Also the increased tolerance of the doughtowards increased mixing time, fermentation time and water content isdescribed. The use of the Carbohydrate oxidase may result in anincreased volume and improved crumb structure and softness of the bakedproduct, as well as an increased strength, stability and reducedstickiness of the dough.

Aims of the Invention

The present invention aims to provide a novel method for improving theshape and width of cuts of bread.

SUMMARY OF THE INVENTION

The present invention concerns a method for the improvement of shape andwidth of cuts of bread during the baking process of bakery productswhich comprises the step of adding a sufficiently effective amount ofCarbohydrate oxidase and/or Pyranose oxidase in said bakery products.

The method of the present invention is in particular suitable for theimprovement of shape and width of cuts of bread during the bakingprocess of bakery products, said baking process comprising forming loafsof a dough, proofing said dough, incising said dough at its top surfaceand baking said dough, and is characterised in that the methodcomprises:

-   -   adding a sufficiently effective amount of carbohydrate oxidase        and/or pyranose oxidase in said dough before proofing,    -   incising said dough at its top surface before or after proofing,        and    -   obtaining a baked product with at its top surface an improved,        wider cut when compared with a reference bread not comprising a        sufficiently effective amount of Carbohydrate oxidase and/or        Pyranose oxidase.

Preferably, the method according to the invention further comprises thestep of adding other conventionally used baking additives orcombinations of baking additives selected from the group consisting ofgluten, oxidantia like vitamin C and Azodicarbonamide, emulsifiers likemono- or diglycerides, diacetyl tartaric acid of monoglycerides, sodiumstearoyl lactylates, sugar esters of fatty acids, lecithin, sugar, salt,fat and/or oil.

Preferably, the method according to the invention is furthercharacterised in that the Carbohydrate oxidase and/or Pyranose oxidaseis added to the ingredients as a dry powder, a granulated oragglomerated powder or a liquid improver.

Preferably, the method according to the invention is furthercharacterised in that the sufficiently effective amount of thecarbohydrate oxidase and/or pyranose oxidase lies between 5 and 500 u/kgflour.

Preferably, the method according to the invention is furthercharacterised in that the width of cuts of the baked product issignificantly larger than compared with a reference bread not comprisinga sufficiently effective amount of Carbohydrate oxidase and/or Pyranoseoxidase.

Advantageously, the method according to the invention is furthercharacterised in that the width of cuts of the baked product is betweenabout 10% and about 400% larger than compared with a reference bread notcomprising a sufficiently effective amount of Carbohydrate oxidaseand/or Pyranose oxidase.

Another aspect of the present invention concerns the use of Carbohydrateoxidase and/or Pyranose oxidase in the improvement of the shape and thewidth of cuts of bread during the baking process of bakery products.

SHORT DESCRIPTION OF THE DRAWINGS

FIG. 1 represents the results of the baking test from example 1.

DETAILED DESCRIPTION OF THE INVENTION

Properties of the Carbohydrate Oxidase:

The oligosaccharide oxidase may be obtained from a strain ofMicrodochium or Acremonium, preferably a strain of M. nivale, morepreferably CBS 100236. WO 99/31990 describes the isolation of the geneencoding the Carbohydrate oxidase from M. nivale CBS 100236 andinsertion into E. coli. and a method for producing a carbohydrateoxidase comprising cultivating the host cell under conditions conductiveto expression of the carbohydrate oxidase and recovering thecarbohydrate oxidase.

A preferred carbohydrate oxidase from M. nivale is capable of oxidizingoligosaccharides having a degree of polymerization (DP) of DP2-DP5, at asubstrate concentration of 0.83 mM at a higher rate than thecorresponding monosaccharide. Thus the enzyme can hydrolyze bothmaltodextrins and cellodextrines wherein monosaccharide units are linkedby alpha-1, 4 or beta-1,4 glucosidic bonds, respectively, at a higherrate than glucose. The carbohydrate oxidase can hydrolyze allcellodextrines having DP2-DP5 equally well and at a level around 10-foldhigher than the monosaccharide glucose. The carbohydrate is preferablystable at pH 5-7. A preferred Carbohydrate oxidase from M. nivale hasoptimum activity around pH=6. At 40° C. it is stable in the pH range4-9, but unstable at pH=3.

The carbohydrate oxidase is preferably stable at 20-45° C. A preferredCarbohydrate oxidase from M. nivale has optimum activity around 40° C.At pH 6 it is stable up to 60° C., but inactivated at 70° C. It has adenaturation temperature of 73° C.

Properties of the Pyranose Oxidase:

The Pyranose oxidases, described in the WO97/22257, are enzymes whichcatalyse the oxidation of several monosaccharides at position C2 withthe release of hydrogen peroxide. Glucose in its Pyranose form is thepreferred substrate. A number of other substrates e.g. furanoses such asxylose can also be oxidized by the enzyme. The Pyranose oxidase isdistinct from glucose oxidase (E.C. 1.1.34) which catalyses theoxidation of glucose at position C1 with the concomitant formation ofgluconic acid.

When Pyranose oxidase is added to dough intended for use in thepreparation of baked products, it may exert an oxidizing effect on doughconstituents and thereby serve, e.g. to improve the strength of thegluten structures in the dough and/or baked products and thereby improvethe strength of the dough in addition to the rheological and handlingproperties of the dough. The oxidizing effect is believed to be achievedwhen the enzyme catalyses the oxidation of monosaccharides present inthe flour or dough. WO97/22257 describes an increased volume andimproved crumb structure and softness of the baked product, as well asan increased strength, stability and reduced stickiness of the dough,thus resulting in improved mach inability when Pyranose oxidase is addedto the dough. The Pyranose oxidase can be from a microbial or fungalorigin.

This invention is related to the effect of the use of Carbohydrateoxidase and Pyranose oxidase, added to a bread improving and/or doughimproving composition, on the oven spring and the shape of the resultingbaked product. An example of a Carbohydrate oxidase is described inpatent WO 99/31990 and an example of a Pyranose oxidase is described inthe patent WO 97/22257. Addition of Carbohydrate oxidase or Pyranoseoxidase to a dough intended for use in preparation of baked productswhich are cut, an incision is made with a sharp razor blade, just beforeor at the end (right before baking) of proofing results in an improvedoven spring, increased width of cuts and improved shape of the finalproduct. Resulting breads are more round than breads prepared withoutoxidases or on addition of Glucose oxidase.

The Carbohydrate oxidase or Pyranose oxidase may be added in acomposition containing other enzymes like amylase, xylanase, lipase,glucose oxidase, lipoxygenase, peroxidase, and protease. Thedough-improving and/or bread improving composition may contain acombination of conventionally used baking additives like: gluten,oxidantia like vitamin C and Azodicarbonamide, emulsifiers like mono- ordiglycerides, diacetyl tartaric acid of monoglycerides, sodium stearoyllactylates, sugar esters of fatty acids, lecithin, sugar and/or salt,fat and/or oil. These mixtures of active ingredients can be diluted byappropriate carriers normally used in baker applications like wheatflour, rye flour, starch, water or oil, to obtain a dosage levelappropriate for mixing in doughs for baking properties. The mixtures canbe in powder, granulated, agglomerated or liquid form.

Carbohydrate oxidase or Pyranose oxidase should be dosed between 5000units and 20000 units for 100 kg of flour.

The effect of addition to the dough of respectively Carbohydrate oxidaseand Pyranose oxidase on the volume and the shape of bread are shown inexamples 1 to 8.

The specifications of the standard non-treated wheat flour used for thebaking trials presented in the following examples are: 12.68% protein,0.58% ash, falling number=235 s, Farinograph A=57.44%, Farinograph B=90,Farinograph C=80.

All breads are lengthwise cut (incised) with a sharp razor blade. Thewidth of cut of the resulting breads is measured as the largest distancebetween the two upstanding edges of the cut after baking.

The standard improver used contains: Fungal alpha amylase (Fungamyl75.000, Novozymes) 0.9 g/100 kg flour, xylanase (Belase B210, Beldem) 3g/100 kg flour, vitamin C 10 g, Datem (Multec Data HP20, Beldem) 300g/100 kg flour. This is an example of the standard improver. Absoluteand relative amounts of additives can vary according to local adaptationto wheat flour and process.

EXAMPLE 1 Comparison of the Effect of Glucose Oxidase, CarbohydrateOxidase and Pyranose Oxidase on the Shape of Overnight Fermented Breads(17 Hours, 20° C.)

Dough was prepared with standard non-treated wheat flour (100), water(54), fresh yeast (Gelka, Belgium) (0.35), salt (2) standard improver(1). Dough was mixed in a (Diosna SP24) spiral mixer for 2 minutes atlow speed and 7 minutes at high speed. Dough temperature after mixing is24° C. Dough pieces of 350 g are rounded and fermented at 25° C. for 20minutes. After moulding (Bertrand, Electrolux Baking) baguette shapeddough pieces are fermented for 17 hours at 20° C., cut lengthwise with 3straight cuts of 2 mm depth and 10 cm length who overlap each other ⅓,per bread and baked in a deck oven (210° C., 30 minutes, steaming: 0.2 Lbefore baking, very little after baking). Volume Width of Dough Shape ofbread (cc) cuts (cm) 1. Reference 1350 5 2. GOX 85 u/kg flour slightlymore round 1550 11 3. PO 200 u/kg flour more round 1825 38 Crust lessbrown 4. CO 200 u/kg flour crust less brown 1875 41

When dosed at 200 units/kg flour and fermented at 20° C. for 17 hoursboth Carbohydrate and Pyranose oxidase improve the form of the bread andthe width of the cuts significantly when compared to addition of Glucoseoxidase.

The result of this are depicted in FIG. 1. The figure shows thereference bread (1), bread comprising 85 u/kg flour GOX (2), breadcomprising 200 u/kg flour PO (3) and bread comprising 200 u/kg flour CO(4). It is clear from the figure that the examples obtained according tothe invention (3 and 4) show a remarkable improvement in the overallappearance of the bread due to the volume increase and the increase inthe width of the cuts 5.

EXAMPLE 2 Comparison of the Effect of Glucose Oxidase and CarbohydrateOxidase and Pyranose Oxidase on the Shape of Overnight Fermented Breads(12 Hours, 27° C.)

Dough was prepared with standard non-treated wheat flour (100), water(54), fresh yeast (Gelka, Belgium) (0.35), salt (2) and standardimprover (1). Dough was mixed in a (Diosna SP24) spiral mixer for 2minutes at low speed and 7 minutes at high speed. Dough temperatureafter mixing is 24° C. Dough pieces of 350 g are rounded and fermentedat 25° C. for 20 minutes. After moulding (Bertrand, Electrolux Baking)baguette shaped dough pieces are fermented for 12 hours at 27° C., cutlengthwise with 3 straight cuts of 2 mm depth and 10 cm length whooverlap each other ⅓, per bread and baked in a deck oven (210° C., 30minutes, steaming: 0.2 L before baking, very little after baking).Volume Width of Dough Shape of bread (cc) cuts (cm) 1. Reference-Datem +2100 6 ADA (3 g/100 kg flour) 2. GOX 85 u/kg flour slightly more round2125 4 3. PO 200 u/kg flour more round 2100 20 Crust less brown 4. CO200 u/kg flour more round 2075 22 crust less brown

When dosed at 200 units/kg flour and fermented at 27° C. for 12 hoursboth Carbohydrate and Pyranose oxidase improve the form of the bread andthe width of the cuts significantly when compared to addition of Glucoseoxidase.

EXAMPLE 3 Effect of Different Dosages of Carbohydrate Oxidase on Widthof Cuts of Overnight Fermented Bread

Dough was prepared with standard non-treated wheat flour (100), water(54), fresh yeast (Gelka, Belgium) (0.35), salt (2) and standardimprover (1). Dough was mixed in a (Diosna SP24) spiral mixer for 2minutes at low speed and 7 minutes at high speed. Dough temperatureafter mixing is 24° C. Dough pieces of 350 g are rounded and fermentedat 25° C. for 20 minutes. After moulding (Bertrand, Electrolux Baking)baguette shaped dough pieces are fermented overnight, cut lengthwisewith 3 straight cuts of 2 mm depth and 10 cm length who overlap eachother ⅓, per bread and baked in a deck oven (210° C., 30 minutes,steaming: 0.2 L before baking, very little after baking). Tests havebeen performed with both non-treated Belgian flour and Argentinianflour.

Specifications of Argentinian flour are: Alveograph P=73 mm H₂O,Alveograph L=78 mm, Alveograph W=197 10E-4 J.

3.1. Belgian Flour, Fermentation: 17 Hours, 20° C. Volume Width of DoughShape of bread (cc) cuts (cm) 1. Reference with Slightly round 1800 18GOX 85 u/kg flour 2. CO 50 u/kg flour round 1750 25 3. CO 100 u/kg flourround 1900 38 4. CO 200 u/kg flour round 1900 37

3.2. Argentinian Flour, no Datem, ADA, Fermentation: 12 h, 27° C. VolumeWidth of Dough Shape of bread (cc) cuts (cm) 1. Reference with GOX 18505 85 u/kg flour − Datem + ADA (3 g/100 kg flour) 2. CO 100 u/kg flourMore round 2000 12 3. CO 200 u/kg flour More round 2000 18

When fermented at 20° C. for 17 hours, the optimum dosage of addedCarbohydrate oxidase to obtain maximum width of cuts is 100 u/kg flour.When fermented at 27° C. for 12 hours the optimum dosage of addedCarbohydrate oxidase to obtain maximum width of cuts is 200 u/kg flour.

EXAMPLE 4 Effect of Different Dosages of Pyranose Oxidase on Width ofCuts of Overnight Fermented Bread

Dough was prepared with standard non-treated wheat flour (100), water(54), fresh yeast (Gelka, Belgium) (0.35), salt (2) and standardimprover (1). Dough was mixed in a (Diosna SP24) spiral mixer for 2minutes at low speed and 7 minutes at high speed. Dough temperatureafter mixing is 24° C. Dough pieces of 350 g are rounded and fermentedat 25° C. for 20 minutes. After moulding (Bertrand, Electrolux Baking)baguette shaped dough pieces are fermented overnight, cut lengthwisewith 3 straight cuts of 2 mm depth and 10 cm length who overlap eachother ⅓, per bread and baked in a deck oven (210° C., 30 minutes,steaming: 0.2 L before, very little after).

Tests have been performed with both non-treated Belgian flour andArgentinian flour. Specifications of Argentinian flour as in example 3.

4.1. Belgian Flour, Fermentation: 17 Hours, 20° C. Volume Width of DoughShape of bread (cc) cuts (cm) 1. Reference with Slightly round 1725 17GOX 85 u/kg flour 2. PO 50 u/kg flour round 1675 12 3. PO 100 u/kg flourround 1775 29 4. PO 200 u/kg flour round 1775 25

4.2. Argentinian Flour, No Emulsifier, ADA, Fermentation: 12 h, 27° C.Volume Width of Dough Shape of bread (cc) cuts (cm) 1. Reference withGOX Slightly round 2050 7 85 u/kg flour − Datem + ADA (3 g/100 kg flour)2. CO 100 u/kg flour round 2250 20 3. CO 200 u/kg flour More round 225026 than 2

The same conclusions as made in example 3 on addition of Carbohydrateoxidase can be made on addition of Pyranose oxidase. When fermented at20° C. for 17 hours the optimum dosage of added Carbohydrate oxidase is100 u/kg flour while for fermentation at 27° C. for 12 hours maximumwidth of cut is measured on addition of 200 u/kg flour.

EXAMPLE 5 Comparison of the Effect of Double Dosage of Glucose Oxidaseand Optimum Dosage of Pyranose Oxidase on Width of Cuts of OvernightFermented Breads

Dough was prepared with standard non-treated wheat flour (100), water(54), fresh yeast (Gelka, Belgium) (0.35), salt (2) and standardimprover. Dough was mixed in a (Diosna SP24) spiral mixer for 2 minutesat low speed and 7 minutes at high speed). Dough temperature aftermixing is 24° C. Dough pieces of 350 g are rounded and fermented at 25°C. for 20 minutes. After shaping baguette shaped dough pieces arefermented for 17 hours at 20° C., cut lengthwise with 3 straight cuts of2 mm depth and 10 cm length who overlap each other ⅓, per bread andbaked in a deck oven (210° C., 30 minutes, steaming: 0.2 L before andvery little after). Volume Width of Dough Shape of bread (cc) cuts(cm) 1. GOX 85 u/kg flour 1925 25 2. GOX 170 u/kg flour 1750 15 3. PO100 u/kg flour more round 1900 37

When doubling the usual dosage of added Glucose oxidase width of cuts isnot increased. The shape and width of cuts of the breads prepared withPyranose oxidase are significantly better than those of the referencebread with Glucose oxidase.

EXAMPLE 6 Effect of Carbohydrate Oxidase, Pyranose Oxidase and GlucoseOxidase on the Shape and the Width of Cuts of Argentinian Breads (DirectProofing and Baking)

Dough was prepared with standard non-treated wheat flour (100), water(54), fresh yeast (Gelka, Belgium) (0.35), salt (2) and standardimprover. Dough was mixed in a (Diosna SP24) spiral mixer for 2 minutesat low speed and 7 minutes at high speed). Dough temperature aftermixing is 24° C. Dough pieces of 350 g are rounded and fermented at 25°C. for 20 minutes. After shaping baguette shaped dough pieces arefermented for 150 minutes at 40° C., cut lengthwise with 3 straight cutsof 2 mm depth and 10 cm length who overlap each other ⅓, per bread andbaked (210° C., 30 minutes, steaming: 0.2 L before, very little after).Volume Width of Dough Shape of bread (cc) cuts (cm) 1. Reference 1025 92. GOX 85 u/kg flour 1125 16 3. GOX 170 u/kg flour Slightly more wide1100 22 4. CO 100 u/kg flour More wide 1075 35 5. CO 200 u/kg flour Morewide 1050 32 6. PO 100 u/kg flour more wide 1050 36 7. PO 200 u/kg flourmore wide 1125 29

Also in direct fermented and baked Argentine type bread addition ofCarbohydrate oxidase or Pyranose oxidase improves the width of cuts ofthe baked product more than addition of Glucose oxidase.

EXAMPLE 7 Effect of Carbohydrate Oxidase and Pyranose Oxidase on Widthof Cuts of Schnittbrötchen

Flour (Weizenmehl type 550) (100), water (56), fresh bakers yeast(Gelka, Belgium) (1), salt (2), standard improver (3) are mixed with aspiral mixer (Diosna SP24) for 2 minutes at low speed and 8 minutes athigh speed. Dough temperature after mixing is 30° C. After 10 minutesbulk fermentation, dough pieces of 1600 g are rounded and divided(Rotamat). Dough pieces of 30 g are rested for 1 minute at 25° C.,moulded (Bertrand, Electrolux Baking), rested for 8 minutes, cutlengthwise (2 cm depth) closed again, turned upside down and fermentedfor 17 hours at 15° C. before baking in a deck oven (16 minutes at 230°C., 0.11 steam before baking and 0.31 steam after baking).

Specifications of the Weizenmehl used are: 11.4% protein, Fallingnumber=310 s, ash content=0.59% Volume Width of Dough Shape of bread(cc) cuts (cm) 1. Reference 1725 40 2. GOX 85 u/kg flour 1650 40 3. PO100 u/kg flour more round 1600 47 4. PO 200 u/kg flour more round 170052 5. CO 100 u/kg flour more round 1625 53 6. CO 200 u/kg flour moreround 1675 57

Width of cut is significantly improved on addition of Carbohydrateoxidase or Pyranose oxidase to the dough while no effect is noticed onaddition of Glucose oxidase.

EXAMPLE 8 Effect of Carbohydrate Oxidase and Pyranose Oxidase on Widthof Cuts of Schnittbrötchen

Flour (Weizenmehl type 550) (100), water (56), fresh bakers yeast(Gelka, Belgium) (1), salt (2), standard improver (3) are mixed with aspiral mixer (Diosna SP24) for 2 minutes at low speed and 8 minutes athigh speed. Dough temperature after mixing is 30° C. After 10 minutesbulk fermentation, dough pieces of 1600 g are rounded and divided(Rotamat). Dough pieces of 30 g are rested for 1 minute at 25° C.,moulded (Bertrand, Electrolux Baking), rested for 8 minutes, cutlengthwise (2 cm depth) closed again, turned upside down and fermentedfor 50 minutes at 32° C. before baking in a deck oven (16 minutes at230° C., 0.11 steam before baking and 0.31 steam after baking). VolumeWidth of cuts Dough (cc) (cm) 1. Reference 2025 27 2. GOX 85 u/kg flour2075 31 3. GOX 170 u/kg flour 2075 34 4. CO 50 u/kg flour 2050 31 5. CO100 u/kg flour 2025 34 6. CO 200 u/kg flour 2025 32 7. PO 50 u/kg flour2000 35 8. PO 100 u/kg flour 2050 36 9. PO 200 u/kg flour 2025 39

When compared to the reference the width of cuts of Schnittbrötchenprepared with addition of Carbohydrate oxidase or Pyranose oxidase isimproved. The effect of Pyranose oxidase is somewhat more pronounced.

1. A method for improvement of shape and width of cuts of bread duringbaking process of bakery products, comprising forming loafs of a dough,proofing said dough, incising said dough at its top surface and bakingsaid dough, wherein said method further comprises: adding a sufficientlyeffective amount of carbohydrate oxidase and/or pyranose oxidase in saiddough before proofing, incising said dough at its top surface before orafter proofing, and obtaining a baked product with at its top surface animproved, wider cut when compared with a reference bread not comprisinga sufficiently effective amount of carbohydrate oxidase and/or pyranoseoxidase.
 2. The method according to claim 1, further comprising the stepof adding other conventionally used baking additives or combinations ofbaking additives selected from the group consisting of gluten, oxidantialike vitamin C and Azodicarbonamide, emulsifiers like mono- ordiglycerides, diacetyl tartaric acid of monoglycerides, sodium stearoyllactylates, sugar esters of fatty acids, lecithin, sugar, salt, fat andoil to the dough before proofing.
 3. The method according to claim 1wherein the Carbohydrate oxidase and/or Pyranose oxidase is added as adry powder, a granulated or agglomerated powder or a liquid improver. 4.The method according to claim 1, wherein the sufficiently effectiveamount of the carbohydrate oxidase and/or pyranose oxidase is between 5and 500 u/kg flour.
 5. The method according to claim 1, wherein thewidth of cuts of the baked product is significantly larger than comparedwith a reference bread not comprising a sufficiently effective amount ofCarbohydrate oxidase and/or Pyranose oxidase.
 6. The method according toclaim 1, wherein the width of cuts of the baked product is between about10% and about 400% larger than compared with the reference bread notcomprising a sufficiently effective amount of Carbohydrate oxidaseand/or Pyranose oxidase.
 7. (canceled)